论文部分内容阅读
目的:观察华蟾素(cinobufacini)对白血病HL-60细胞的增殖抑制作用和诱导凋亡作用的机制。方法:以HL-60细胞为研究对象,采用MTT法观察细胞增殖作用;Annexin V-FITC/PI双染和吖啶橙/溴乙锭(AO/EB)荧光染色法检测细胞凋亡;罗丹明染色法检测线粒体膜电位;分光光度法检测Caspase-3活性。结果:在0.78～6.25μg·m~(-1),华蟾素能明显的抑制HL-60细胞的增殖;华蟾素作用24h后,Annexin V阳性的细胞从7.81%增加至66.02%,而且在荧光显微镜下可以明显观察到凋亡细胞;线粒体膜电位下降和Caspase-3活性升高。结论:华蟾素能够抑制HL-60细胞增殖,这种作用与其诱导细胞凋亡破坏线粒体功能和激活Caspase-3有关。
OBJECTIVE: To observe the inhibitory effect of cinobufacini on leukemia HL-60 cells and the mechanism of apoptosis induction. METHODS: HL-60 cells were used as research objects. MTT assay was used to observe cell proliferation. Annexin V-FITC/PI double staining and acridine orange/ethidium bromide (AO/EB) fluorescent staining were used to detect apoptosis. Rhodamine Mitochondrial membrane potential was detected by staining; Caspase-3 activity was measured by spectrophotometry. RESULTS: In the range of 0.78-6.25 μg·m-1, Cinobufacin significantly inhibited the proliferation of HL-60 cells. After 24 hours of cinobufacin treatment, the number of Annexin V-positive cells increased from 7.81% to 66.02%. Apoptotic cells were clearly observed under fluorescence microscope; mitochondrial membrane potential decreased and Caspase-3 activity increased. CONCLUSION: Cinobufacini can inhibit the proliferation of HL-60 cells, which is related to the induction of apoptosis and the destruction of mitochondrial function and activation of Caspase-3.